Evidence on differential role for alpha 1 and alpha 2 subtypes of AP-2 adaptin in the central nervous system.

Two subtypes of alpha (α)subunits, α1and α2, belonging to AP-2 complex have been described in the central nervous system (CNS). The specific role of each subtype is still unclear. In this study, we evaluated the expression and interaction with cell membranes of both subtypes in the postnatal developing cerebral cortex and cerebellum in two rat strains that display distinct developmental features. We observed that α2 displays higher variations than α1 during development, and at lesser extent in the rats with delayed rate of development. Additionally, by in vitro binding assays we evaluated the interaction of α subunits with bovine brain membranes. Both subtypes displayed clear differences in their performance, maximum binding of α1 was higher and α2 reached it faster than α1. In addition, both subtypes displayed different binding to membranes when bivalent cations or nucleotides were added. We conclude that both subtypes interact differently with membranes and that they may play different roles in clathrin-mediated endocytosis in the CNS.

Anestesia para cesárea en el síndrome de Gitelman / Anesthesia for cesarean delivery in Gitelman syndrome

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Effect of cholesterol-5 alpha,6 alpha-epoxide supplementation to cultured cardiomyocytes.

In order to evaluate the effect of one of the main oxysterols derived from cholesterol oxidation, cholesterol-5 alpha,6 alpha-epoxide (epox), on cardiac cells, we have supplemented the culture medium of neonatal rat cardiomyocytes with scalar concentrations of epox (0.1-100 microM). While 0.1 microM epox supplementation was ineffective, epox supplementation in the range 1-100 microM determined a reduction in cellular protein level, without affecting cell viability, and a dose-dependent epox incorporation into cardiomyocyte lipids. Furthermore, in the same concentration range of epox supplementation, a gas chromatographic peak unambiguously identified by gas chromatography-mass spectrometry as cholestane-3 beta,5 alpha,6 beta-triol, an hydrolytic metabolite of epox, was detected. The mechanism of cytotoxicity of epox to cardiomyocytes could be due to the insertion of epox itself into cellular lipids, and to its metabolization to the more toxic triol.

Real-time monitoring of electrically stimulated norepinephrine release in rat thalamus: I. Resolution of transmitter and metabolite signal components.

Electrical stimulation of an ascending path of the locus ceruleus-norepinephrine system was used to elicit release of norepinephrine at noradrenergic terminal fields of the rat thalamus. Overflow into the extracellular fluid space was measured by fast in vivo chronoamperometry. At pretreated carbon fibers, the electrochemical signal consists of a sharp peak of approximately 20-30 s duration followed by a slower, plateau-like decay to baseline. The peak, characterized by a variety of pharmacological manipulations and dialysis perfusion, is primarily due to norepinephrine. The plateau was shown to correspond to metabolite efflux of 3,4-dihydroxy-phenylacetic acid. By varying the degree of electrochemical pretreatment, the response time and sensitivity of the fibers can be tuned to follow the entire signal or to select the separate components for detailed evaluation. This approach can be used to provide new information on the spatial and temporal characteristics of stimulated neurotransmitter release.

Real-time monitoring of electrically stimulated norepinephrine release in rat thalamus: II. Modeling of release and reuptake characteristics of stimulated norepinephrine overflow.

As in the preceding study, electrical stimulation was used to effect release overflow of norepinephrine in the rat thalamus. Using a weak electrochemical pretreatment of a carbon fiber electrode, it was possible to "tune in" the electrochemical response signal for norepinephrine without metabolite interference. This reasonably selective signal was then used to study the degradation of norepinephrine release ability caused by prolonged stimulation. Further, the signals were modeled by the method used successfully for stimulated dopamine overflow, providing hitherto unavailable information on the temporal and spatial characteristics of norepinephrine release overflow. Pertinent comparisons between the release characteristics of the dopamine and norepinephrine systems show that the half-life for norepinephrine in the extracellular fluid space is approximately 1 s in thalamus compared with 33 ms for dopamine in caudate.

[Saturated fatty acids determination by GLC after TLC separation (author's transl)]. / Determinazione degli acidi grassi saturi mediante GLC previa separazione su strato sottile

The determination of fatty acid composition of oils and fats by conventional GLC on polar columns gives satisfactory results for most of the applications. Higher precision is, however, desirable when the content of minor components is of interest. A typical example is the quantitative determination of the percentage of peanut oil in a seed oil mixture, on the basis of the content of lignoceric acid (n-C21). Lignoceric acid, wich is containen in peanut oil in the amount of 1-2%, shows, on polar columns, a very long retention time and, owing to its low percentage, flat and non well measurable peaks. A method was therefore developed which allows an accurate determination of all saturated fatty acids, particulary of those with high molecular weight. The method is based on the separation of satured fatty acids by argentation TLC followed by GLC determination on non polar columns.

Hydroxy fatty acids from cerebrosides of the central nervous system: GLC determination and mass spectrometric identification.

A method has been developed for GLC determination of hydroxy fatty acids (HFA) from cerebrosides of the central nervous system.Identification of HFA present in complex biological mixtures was achieved by the use of the combination gas chromatography- mass spectrometry. GLC separations and mass spectrometric determinations were carried out on the trimethylsilyl derivatives of HFA, which show excellent GLC characteristics and favorable cleavages to determine the position of OH groups on the aliphatic chain. The mass spectra of these derivatives present some unusual rearrangement ions, whose composition has been studied with the aid of deuterium labeled analogs.

Isolation of geranyl geraniol from the unsaponifiable fraction of linseed oil.

From the unsaponifiable fraction (63 g) of linseed oil (25 kg), two terpenic alcohols were isolated by alumina column, thin-layer, and gas-liquid chromatography. They were identified as phytol and geranyl geraniol (a precursor of bi- and tricyclic diterpenes) by infrared and nuclear magnetic resonance spectroscopy, ozonolysis, and mass spectrometry.